Project 2: Serologic and molecular determinants of COVID-19 severity and immune protection

项目2摘要 项目2. COVID-19严重程度和免疫保护的血清学和分子决定因素。 尚未解决的科学问题仍然是关于SARS-CoV-2血清学的类型、动力学和持续时间如何影响SARS-CoV-2的感染。 反应与患者年龄、疾病严重程度、保护性免疫力和传播至近距离接触的风险相关。 联系人.我们的总体目标是利用注释良好的STOP-COVID数据集和纵向样本 从首个应答者队列（再次暴露的高风险）和家庭接触者（项目1，核心B）中收集， 深入了解持久和保护性血清学的特定宿主和病毒因素 应答我们的三个高度互动的目标包括：确定病毒中和（VN）抗体的差异 同种型及其靶病毒蛋白/肽/表位;评估针对疾病的交叉保护性免疫 通过病毒宿主转录组分析， SARS-CoV滴度或序列以及粘膜部位或血液中的初始免疫应答与疾病相关 结果和上述血清学反应。目的1：定义免疫球蛋白同种型、滴度、靶蛋白和 SARS-CoV-2特异性抗体的病毒中和活性及其与疾病严重程度和 保护：我们将利用VN检测（VNA）和荧光素酶免疫沉淀检测来鉴定VN抗体 同种型特异性及其免疫显性和独特的靶病毒蛋白/肽/表位 疾病的严重性和保护。预期结果包括描述时间抗体同种型的数据 和病毒靶点，以完善与保护性免疫一致的血清学检测方案。目标2：确定 普通感冒冠状病毒（CCCoV）和SARS-CoV-2特异性抗体对COVID-19保护的影响， 疾病的严重程度在一个高暴露风险队列的第一反应者和他们的家庭接触。诊断 血清样本将用于针对独特和保守的CCCoV的创新ELISA平台 肽和Spike假型化CCCoV以评估CCCoV抗体应答。预期成果包括 关于既存CCCoV抗体是否与COVID-19临床症状、严重程度或 SARS-CoV-2感染的菌株特异性（目标3）或再感染风险。目标3。相关的粘膜模式和 血清学免疫应答与病毒和宿主遗传学。我们将利用一种新的靶向宿主-SARS-CoV-2 RNA测序分析（CoV-DXVX），以评估SARS-CoV-2 S1中的病毒滴度或功能改变， S2、N蛋白或其他ORF影响疾病的表现、进展和结果。宿主反应 通过粘膜或血液免疫基因诱导的特征和强度来测量， 这些血清学应答的持续时间、类型和保护能力以及与上述血清学应答的关系。一旦疫苗 部署到我们的第一反应者队列，所有3个目标将重点分析上述血清学反应如何 在接种SARS-CoV-2血清阴性和血清阳性的患者中， 与自然感染者相比。

Project 2 Summary PROJECT 2. Serologic and molecular determinants of COVID-19 severity and immune protection. Unresolved scientific questions remain about how the type, kinetics and duration of SARS-CoV-2 serologic responses correlate with patient age, disease severity, protective immunity, and risk of spread to close contacts. Our overall goal is to utilize the well-annotated STOP-COVID data sets and longitudinal samples collected from the first-responder cohort (at high-risk for re-exposure) and family contacts (Project 1, Core B) to advance understanding of the specific host and viral factors that underlie long-lasting and protective serologic responses. Our three highly interactive aims include: to identify differences in virus neutralizing (VN) antibody isotypes and their target viral proteins/peptides/epitopes; to assess cross-protective immunity versus disease exacerbation by antibodies to common cold CoVs; and to probe, by virus-host transcriptome analysis, how SARS-CoV titer or sequence and initial immune response at mucosal sites or blood interrelate with disease outcomes and the above serologic responses. Aim 1: Define immunoglobulin isotypes, titer, target proteins and viral neutralization activity of SARS-CoV-2 specific Abs and their correlations with disease severity and protection: We will utilize VN assays (VNA) and a luciferase immunoprecipitation assay to identify VN antibody isotype specificities and their immunodominant and unique target viral proteins/peptides/epitopes relevant to disease severity and protection. Expected outcomes include data that delineate the temporal antibody isotypes and viral targets to refine protocols for serologic testing that align with protective immunity. Aim 2: Determine the impact of common cold CoVs (CCCoV) and SARS-CoV-2 specific antibodies on COVID-19 protection and disease severity in a high-exposure risk cohort of first responders and their household contacts. Diagnostic serum samples will be used in innovative ELISA platforms directed against unique and conserved CCCoV peptides, and Spike pseudotyped CCCoV to assess CCCoV antibody responses. Expected outcomes include original data on whether pre-existing CCCoV antibodies relate to COVID-19 clinical symptoms, severity or strain specificity of SARS-CoV-2 infection (Aim 3), or re-infection risk. Aim 3. Correlate patterns of mucosal and serologic immune responses with virus and host genetics. We will utilize a novel targeted host-SARS-CoV-2 RNA sequencing assay (CoV-DXVX) to assess whether virus titer or functional alterations in SARS-CoV-2 S1, S2, N proteins or other ORFs impact disease presentation, progression and outcome. Host response measured by the profile and strength of mucosal or blood immune gene induction will be correlated with the duration, type and protective ability of these serologic responses and with the ones above. Once vaccines are deployed to our first-responder cohort, all 3 aims will pivot to analyze how the above serologic response profiles and host immune gene induction differ in vaccinated SARS-CoV-2 seronegative and seropositive individuals compared with the naturally-infected individuals.